Purification and Cloning of Carp Nephrosin, a Secreted Zinc Endopeptidase of the Astacin Family
Resource
THE JOURNAL OF BIOLOGICAL CHEMISTRY 272 (21): 13772-13778
Journal
THE JOURNAL OF BIOLOGICAL CHEMISTRY
Journal Volume
272
Journal Issue
21
Pages
13772-13778
Date Issued
1997
Date
1997
Author(s)
Abstract
We have purified a secreted proteinase of 23 kDa from carp head kidney by sequential column chromatography on a Reactive Blue 72-agarose dye affinity column and an FPLC Mono-P column. The secretion of this proteinase from carp head kidney can be stimulated by high concentrations of potassium. Since the carp proteinase is present mainly in the head kidney, kidney, and spleen (all of which are lymphohematopoietic organs), it is named nephrosin. The carp nephrosin is most sensitive to metal chelators, but not to inhibitors specific for other classes of proteinases. A cDNA clone has been isolated from a carp head kidney cDNA library by immunoscreening with a polyclonal antiserum raised against purified nephrosin. The cloned cDNA is 1086 base pairs in length and has an open reading frame encoding a protein of 273 amino acids, including a 19-amino acid signal peptide and 56-amino acid propeptide. The deduced amino acid sequence shows moderate levels of identity to medaka HCE1 (52.5%), medaka LCE (50.7%), crayfish astacin (33.2%), murine meprin-α (34%), and murine meprin-β (33.5%), all members of the astacin family of zinc endopeptidases. Nephrosin is the first member of the astacin family found in lymphohematopoietic tissues.
Other Subjects
proteinase; animal tissue; article; carp; column chromatography; dna library; enzyme purification; extracellular matrix; hematopoietic cell; molecular cloning; nonhuman; noradrenalin release; open reading frame; priority journal; protein secretion; proteinase inhibition; teleost; Adrenal Cortex; Amino Acid Sequence; Animals; Base Sequence; Blotting, Northern; Carps; Chromaffin Cells; Cloning, Molecular; DNA, Complementary; Kidney; Metalloendopeptidases; Molecular Sequence Data; Open Reading Frames; Perfusion; Potassium; Protease Inhibitors; Sequence Alignment; Substrate Specificity; Tissue Distribution; Animalia; Astacoidea; Cyprinus carpio; Murinae; Teleostei
Type
journal article
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